Ab417 binds to L1CAM-expressing ICC cells and reduces L1CAM levels at the membrane.

<p>(A) Confocal microscopic analysis of Ab417 internalization. SCK-L1 cells were preincubated with Ab417 (10 μg/mL) for the indicated times. Membrane L1CAM-bound Ab417 was detected using anti-Human IgG(H+L) Cross Adsorbed DyLight 594 (red), and internalized Ab417 was detected using anti-human IgG (Fc-specific)-FITC (green). (B) Choi-CK cells were incubated with Ab417 (10 μg/mL) for the indicated times, and the membrane L1CAM was detected by western blot analysis using A10-A3 and pan-cadherin as a loading control. Control (c) indicates the cells that were incubated for 6 h without Ab417. (C) Confocal microscopic images of L1CAM staining of PBS (up)- or Ab417 (down)-treated Choi-CK tumors. The images were taken at x400 magnification. L1CAM index (D) are represented as a percentage of positively-stained area comparing to total area.</p

Anti-tumor efficacy of gemcitabine or cisplatin in a Choi-CK xenograft model.

<p>Drug at indicated dose was <i>i</i>.<i>p</i>. injected twice a week for 3 or 4 weeks, and tumor volume (A) and tumor weight (B) were determined. Each point indicates the mean ± s.d. <i>p</i> < 0.05 (*) and <i>p</i> < 0.01 (**), significant difference from the saline-treated group by Dunnett’s <i>t</i>-test. <i>p</i> < 0.05 (^#) and <i>p</i> < 0.01 (^##), significant difference from the saline group by Steel’s test.</p

Anti-tumor efficacy of combined treatment with Ab417 and gemcitabine or cisplatin in a Choi-CK model.

<p>Tumor volume (A and C) and tumor weight (B and D) are represented. Each point indicates the mean ± s.d (<i>n</i> = 8). <i>p</i> < 0.05 (*) and <i>p</i> < 0.01 (**), significant difference from the saline group by Dunnett’s <i>t</i>-test. <i>p</i> < 0.05 (^#) and <i>p</i> < 0.01 (^##), significant difference from the saline group by Steel’s test.</p

Anti-tumor efficacy of Ab417 in a Choi-CK xenograft model.

<p>Ab417 (10 mg/kg) or the isotype control (hFc, 3.3 mg/kg) was <i>i</i>.<i>v</i>. injected twice a week into nude mice bearing Choi-CK or TFK-1 xenografts (<i>n</i> = 8). Tumor volume (A), tumor weight (B), and body weight (C) were represented. Each point indicates the mean ± s.d. <i>p</i> < 0.05 (*) and <i>p</i> < 0.01 (**), significant difference from the isotype control group by Dunnett’s <i>t</i>-test. <i>p</i> < 0.05 (^#), significant difference from the isotype control group by Steel’s test. (D) Tumor sections were stained with an anti-human Ki-67 antibody. Ki-67 index are represented as a percentage of positively-stained area comparing to total area.</p